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Western blot analysis of Ubiquitin on different lysates with Rabbit anti-Ubiquitin antibody (ER31212) at 1/500 dilution.
Lane 1: MCF-7 cell lysate
Lane 2: MCF-7 cell lysate treated with 50 µM MG132 for 1.5h
Lysates/proteins at 10 µg/Lane.
Predicted band size: 8 kDa
Exposure time: 2 minutes;
15% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER31212) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of Ubiquitin on different cell lysates using anti-Ubiquitin antibody at 1/1,000 dilution.
Positive control:
Lane 1: 293T cell lysate
Lane 2: HepG2 cell lysate
Lane 3: Hela cell lysate
Lane 4: MCF-7 cell lysate
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ICC staining Ubiquitin in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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ICC staining Ubiquitin in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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ICC staining Ubiquitin in MCF-7 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER31212) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER31212) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunocytochemistry analysis of NIH/3T3 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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Immunocytochemistry analysis of PC-12 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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